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Methods to Study Epithelial Transport Protein Function and Expression in Native Intestine and Caco-2 Cells Grown in 3D
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Serotonin modifies cytoskeleton and brush-border membrane architecture in human intestinal epithelial cells

Ravinder K Gill1, Le Shen, Jerrold R Turner

  • 1Dept. of Medicine, Univ. of Illinois at Chicago, Medical Research Service, Jesse Brown VA Medical Center, 820 S. Damen Ave., Chicago, IL 60612, USA. rgill@uic.edu

American Journal of Physiology. Gastrointestinal and Liver Physiology
|August 2, 2008
PubMed

Insights

Serotonin (5-hydroxytryptamine) alters intestinal cell structure by affecting the actin cytoskeleton via 5-HT3/4 receptors. This impacts ion transporter function, potentially contributing to diarrheal diseases.

Area of Science:

  • Gastroenterology
  • Cell Biology
  • Molecular Pharmacology

Background:

  • Serotonin (5-hydroxytryptamine, 5-HT) regulates gastrointestinal functions.
  • Previous studies showed 5-HT inhibits Na+/H+ exchangers (NHE) and Cl-/OH- exchangers in Caco-2 cells.
  • Cytoskeletal integrity influences ion transporter regulation, including NHE3.

Purpose of the Study:

  • To investigate if 5-HT modifies the actin cytoskeleton and brush-border membrane architecture.
  • To identify the signaling pathways involved in 5-HT-induced cellular changes.

Main Methods:

  • Ultrastructural analysis of Caco-2 cells treated with 5-HT.
  • Fluorescence microscopy to examine actin cytoskeleton organization.
  • In vitro assays to assess protein interactions.

Main Results:

  • 5-HT treatment induced apical membrane doming and microvilli shortening.
  • 5-HT promoted basal stress fibers and cortical actin via 5-HT3 and 5-HT4 receptors.
  • These changes were partially dependent on intracellular Ca2+ and PKCalpha.
  • PKCalpha interaction with actin increased upon 5-HT treatment.

Conclusions:

  • 5-HT signaling through 5-HT3/4 receptors regulates the actin cytoskeleton.
  • Ca2+ and PKCalpha mediate 5-HT's effects on cytoskeletal dynamics.
  • This cytoskeletal modulation influences ion transporters, relevant to diarrheal conditions.

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